CLAffinity: A Software Tool for Identification of Optimum Ligand Affinity for Competition-Based Primary Screens.

ABSTRACT: A simplistic assumption in setting up a competition assay is that a low affinity labeled ligand can be more easily displaced from a target protein than a high affinity ligand, which in turn produces a more sensitive assay. An often-cited paper correctly rallies against this assumption and recommends the use of the highest affinity ligand available for experiments aiming to determine competitive inhibitor affinities. However, we have noted this advice being applied incorrectly to competition-based primary screens where the goal is optimum assay sensitivity, enabling a clear yes/no binding determination for even low affinity interactions. The published advice only applies to secondary, confirmatory assays intended for accurate affinity determination of primary screening hits. We demonstrate

SUBMITTER: Shave S 

PROVIDER: S-EPMC9131445 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature